兔角膜缘干细胞体外无血清培养及生物学特性初步研究  

In vitro serum-free culture and biological characteristics preliminary researchof rabbit limbal stem cells

作　　者：黄晓梅[1] 马蕾[2] 赵越超[1] 杨燕妮[1] 赵博[1] 

机构地区：[1]山西医科大学,太原030001 [2]山西医科大学第二临床医学院眼科

出　　处：《中国医疗前沿》2011年第19期43-44,22,共3页China Healthcare Innovation

基　　金：国家自然科学基金(编号:30973248);山西省回国留学人员科研资助基金(编号:200860);山西省留学人员科技择优项目基金(编号:200856)

摘　　要：目的探讨兔角膜缘干细胞体外无血清培养方法及生物学特性并与传统的10%血清培养方法进行比较。方法获取兔角膜缘组织,酶消化法制成单细胞悬液,分含B27的无血清组和传统方法的10%血清组两组进行培养,观察两组细胞的生长情况,CCK-8检测其克隆增殖能力,细胞免疫化学染色方法对两组细胞进行鉴定。结果较传统10%血清培养组,无血清培养组培养的兔角膜缘干细胞表现出相似的生长特性及增殖能力,细胞免疫化学染色无血清组⊿Np63阳性细胞较10%血清组明显多,角蛋白K3(AE5)阳性细胞10%血清组较多。结论用含B27的无血清培养基可以培养获得角膜缘干细胞,减少了血清中不明成分对干细胞的影响,为角膜缘干细胞的体外培养、扩增及纯化提供了新的选择。Objective To investigate ex vivo serum-free culture method of rabbit limbal stem cells and compared with 10% serum culture method.Method Limbal epithelium harvested from rabbits were enzymatically cleft into single cell suspension and seeded in serum-free and 10% serum medium.The growth status of LSCs was observed,the proliferative capacity was assessed by CCK-8,Keratin3 and ⊿Np63 expression were analyzed by immunostaining.Results The cells cultured in the two different mediums show similar growth characteristics and proliferation capacity,cells immunostained with ⊿Np63 was more in serum-free medium and Keratin 3 more in 10% serum medium.Conclusion LSCs was successfully cultured in serum-free medium,it can reduce the influence of unknown components in serum on LSCs,and provides new options for LSCs in vitro culture,amplification and purification.

关 键 词：干细胞 角膜缘 B27 ⊿Np63 细胞培养 

分 类 号：R329.3[医药卫生—人体解剖和组织胚胎学]